Host: UL; Supervisor: Polona Žnidaršič Plazl
Objectives
-
Screening of immobilisation methods suitable for retaining various biocatalysts within the MBR. (M7-M25)
-
Cooperation in development of magnetic-field supported MBR. (M10-M22)
-
Biocatalyst screening using enzymatic or whole-cell MBRs. (M10-M34)
-
Characterisation of enzymatic reactions, performed within various MBRs, including estimation of kinetic parameters. (M18-M42)
-
Intensification of enzymatic reactions with product removal - either in situ or with downstream process integration. (M20 – M42)
Tasks and methodology
ESR7 will screen various novel immobilization methods (e.g. the use of nanotubes, magnetic nanoparticles and tagged fused proteins) in order to achieve high enzyme load with high activity within a MBR. In cooperation with partner iX-factory ESR7 will develop a magnetic-field supported MBR, where immobilization of cells and biomolecules using magnetic nanoparticles, obtained from nanoMyP, could be retained (WP6). Such MBRs will be used for the rapid evaluation of kinetic parameters of single enzyme variants. The immobilization efficiency, as well as biocatalyst activity and stability will be tested for Saccharomyces cerevisiae in L-malic acid production, as well as with purified fumarase. Different biotransformations with lipases and laccase will be tested within various solvent systems including ionic liquids as green solvents. Monitoring methods will also be implemented into a microfluidic motherboard in order to enable process description and model-based optimization. ESR7 will perform investigations on the comparative screening of different biocatalytic reactions with enzymes and cells at the reactor wall and with packed bed or fluidized bed MBR, as well as in batch, fed batch and continuous mode. (WP7) Furthermore, Micronit’s and iX-factory´s reactor technology will be used to establish and model enzymatic reactions and phase separations to recover product and thereby intensify the process by integration with downstream processing.
Results
-
Established non-covalent immobilisation methods suitable for IMERs using magnetic nanoparticles
-
Microfluidic platform for chemical and enzymatic reactions, amenable for different immobilization methods
-
Evaluation of kinetic parameters for free and immobilized enzymes.
-
Evaluation of immobilization methods suitable for wide range of biocatalysts and for multi-enzyme and chemo-enzyme process conditions completed.
-
Different biotransformations, e.g. lipase-catalyzed isoamyl acetate production in one and multi-phase systems including ionic liquids characterized.
-
Biocatalyst screening results using developed reversible immobilization technique completed.
-
Magnetic nano-sensor particles tested in biocatalysis (WP7).